Imperial/Wet Lab/Lab Notebook/2007-09-14

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__NOTOC__
= 14 September 2007 =
= 14 September 2007 =
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==Restriction Digest of Biobricks==
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#4 DNA samples were digested by SpeI and PstI
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#* Samples 5, 7, 24
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#1 DNA sample wasdigested by XbaI and PstI
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#* Samples 23
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#4 DNA samples were digested by EcoRI
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#* Samples 13, 25, 27b-e, T7, Kirsten's parts
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#4 μl of DNA was digested by 1 μl of enzymes at 37&deg;C for 1 hour<br>Exception: Sample 23 had 6 μl DNA and 2 μl enzymes
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Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Restriction_Digest | Restriction Digest] in the general protocols page
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==Agarose Gel Electrophoresis of Biobricks==
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#Checked 9 Biobricks and 13 Digests on 1% agarose gel
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[[Image:ICGEMS Gel 14 9.png|900px|]]
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Conclusions:
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*Part 27 is confirmed to be faulty
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*T7 maxi-prep was not done properly
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*Kirsten's parts were most likely not there...
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Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis | Electrophoresis] in the general protocols page
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==Gel Purification of Biobricks==
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#Gel was melted and run through column
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#Purified DNA was eluted with ddH<sub>2</sub>O
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<font size=-2>
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*5. BBa_R0040 [ptet] (~2100 bp fragment)
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*7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
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*23. BBa_I13504 [GFP] (~800 bp fragment)
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*24. BBa_R0062 [pLux] (~2100 bp fragment)
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Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Gel_Purification | Gel Purification] in the general protocols page
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Latest revision as of 19:01, 26 October 2007

14 September 2007

Restriction Digest of Biobricks

  1. 4 DNA samples were digested by SpeI and PstI
    • Samples 5, 7, 24
  2. 1 DNA sample wasdigested by XbaI and PstI
    • Samples 23
  3. 4 DNA samples were digested by EcoRI
    • Samples 13, 25, 27b-e, T7, Kirsten's parts
  4. 4 μl of DNA was digested by 1 μl of enzymes at 37°C for 1 hour
    Exception: Sample 23 had 6 μl DNA and 2 μl enzymes

Protocols can be found at | Restriction Digest in the general protocols page

Agarose Gel Electrophoresis of Biobricks

  1. Checked 9 Biobricks and 13 Digests on 1% agarose gel

ICGEMS Gel 14 9.png

Conclusions:

  • Part 27 is confirmed to be faulty
  • T7 maxi-prep was not done properly
  • Kirsten's parts were most likely not there...

Protocols can be found at | Electrophoresis in the general protocols page

Gel Purification of Biobricks

  1. Gel was melted and run through column
  2. Purified DNA was eluted with ddH2O

  • 5. BBa_R0040 [ptet] (~2100 bp fragment)
  • 7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
  • 23. BBa_I13504 [GFP] (~800 bp fragment)
  • 24. BBa_R0062 [pLux] (~2100 bp fragment)

Protocols can be found at | Gel Purification in the general protocols page


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